Substrate Reagent Pack (8 vials Color A/B) (DY999): R&D Systems

INTENDED USE & DESCRIPTION

Substrate Reagent Pack contains 8 vials of Color Reagent A (stabilized hydrogen peroxide, Part # 895000) and 8 vials of Color Reagent B (stabilized tetramethylbenzidine, Part # 895001), both at 12 mL/vial, producing color development when contacting HRP-conjugated or biotinylated, streptavidin-HRP bound detection antibodies.   

STORAGE & STABILITY

Store unopened at 2-8 °C. Stable for up to 30 days after opening when stored at 2-8 °C.

REAGENT PREPARATION

Warm to room temperature before use. Color Reagent A and Color Reagent B should be mixed together in equal volumes within 15 minutes of use. Protect from light. Add 100 µL of the mixture to each well and incubate at room temperature for 20 minutes while protecting from light. Add Stop Solution (Catalog # DY994) and read the microplate.

TECHNICAL HINTS & LIMITATIONS OF THE PROCEDURE 

  • Caution: Color Reagent B may cause skin, eye, and respiratory irritation. Avoid breathing fumes. Wear protective gloves, clothing, and eye and face protection. Wash hands thoroughly after handling. Please refer to the MSDS on our website prior to use. 
Preparation and Storage
  • Stability & Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

55 Citations: Showing 1 - 10
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  1. Restoring GM1 ganglioside expression ameliorates axonal outgrowth inhibition and cognitive impairments induced by blast traumatic brain injury
    Authors: V Rubovitch, Y Zilberstei, J Chapman, S Schreiber, CG Pick
    Sci Rep, 2017;7(0):41269. 2017
  2. Neuroprotection by Caffeine in Hyperoxia-Induced Neonatal Brain Injury
    Authors: S Endesfelde, U Weichelt, E Strauá, A Schl”r, M Sifringer, T Scheuer, C Bhrer, T Schmitz
    Int J Mol Sci, 2017;18(1):. 2017
  3. Marked elevation in plasma osteoprotegerin constitutes an early and consistent feature of cerebral malaria
    Thromb. Haemost., 2016;115(4):773-80. 2016
  4. Regulation of C3 Activation by the Alternative Complement Pathway in the Mouse Retina
    PLoS ONE, 2016;11(8):e0161898. 2016
  5. The Robo4 cytoplasmic domain is dispensable for vascular permeability and neovascularization
    Nat Commun, 2016;7(0):13517. 2016
  6. Mechanism of the antigen-independent cytokinergic SPE-7 IgE activation of human mast cells in vitro.
    Authors: Bax H, Bowen H, Dodev T, Sutton B, Gould H
    Sci Rep, 2015;5(0):9538. 2015
  7. Bone marrow-derived macrophages from aged rats are more responsive to inflammatory stimuli.
    Authors: Barrett J, Costello D, O'Sullivan J, Cowley T, Lynch M
    J Neuroinflammation, 2015;12(0):67. 2015
  8. An automated method for determining the cytoadhesion of Plasmodium falciparum-infected erythrocytes to immobilized cells.
    Authors: Hempel C, Boisen I, Efunshile A, Kurtzhals J, Staalso T
    Malar J, 2015;14(0):112. 2015
  9. Upregulated interleukin-21 receptor on B cells associated with the downregulation of IgE in patients with allergic rhinitis.
    Authors: Jen, Hsiao-Yu, Yang, Yao-Hsu, Chiang, Bor-Luen, Chuang, Ya-Hui
    J Interferon Cytokine Res, 2015;35(1):42-8. 2015
  10. Innate immunity drives the initiation of a murine model of primary biliary cirrhosis.
    Authors: Chang C, Chen Y, Zhang W, Leung P, Gershwin M, Chuang Y
    PLoS ONE, 2015;10(3):e0121320. 2015
  11. Macrophage Migration Inhibitory Factor in Acute Adipose Tissue Inflammation.
    Authors: Kim B, Rongisch R, Hager S, Grieb G, Nourbakhsh M, Rennekampff H, Bucala R, Bernhagen J, Pallua N
    PLoS ONE, 2015;10(9):e0137366. 2015
  12. Interleukin-22 is increased in multiple sclerosis patients and targets astrocytes.
    Authors: Perriard G, Mathias A, Enz L, Canales M, Schluep M, Gentner M, Schaeren-Wiemers N, Du Pasquier R
    J Neuroinflammation, 2015;12(0):119. 2015
  13. A C-terminal HSP90 inhibitor restores glucocorticoid sensitivity and relieves a mouse allograft model of Cushing disease.
    Authors: Riebold M, Kozany C, Freiburger L, Sattler M, Buchfelder M, Hausch F, Stalla G, Paez-Pereda M
    Nat Med, 2015;21(3):276-80. 2015
  14. Non-Human Primates Harbor Diverse Mammalian and Avian Astroviruses Including Those Associated with Human Infections.
    Authors: Karlsson E, Small C, Freiden P, Feeroz M, Matsen F, San S, Hasan M, Wang D, Jones-Engel L, Schultz-Cherry S
    PLoS Pathog, 2015;11(11):e1005225. 2015
  15. Early decline in serum phospho-CSE1L levels in vemurafenib/sunitinib-treated melanoma and sorafenib/lapatinib-treated colorectal tumor xenografts.
    Authors: Lee W, Shen S, Shih Y, Chou C, Tseng J, Chin S, Liu K, Chen Y, Jiang M
    J Transl Med, 2015;13(0):191. 2015
  16. Crucial role of IL1beta and C3a in the in vitro-response of multipotent mesenchymal stromal cells to inflammatory mediators of polytrauma.
    Authors: Hengartner N, Fiedler J, Schrezenmeier H, Huber-Lang M, Brenner R
    PLoS ONE, 2015;10(1):e0116772. 2015
  17. Hepatocytes induce Foxp3(+) regulatory T cells by Notch signaling.
    Authors: Burghardt S, Claass B, Erhardt A, Karimi K, Tiegs G
    J Leukoc Biol, 2014;96(4):571-7. 2014
  18. Modulation of endotoxicity of Shigella generalized modules for membrane antigens (GMMA) by genetic lipid A modifications: relative activation of TLR4 and TLR2 pathways in different mutants.
    Authors: Rossi O, Pesce I, Giannelli C, Aprea S, Caboni M, Citiulo F, Valentini S, Ferlenghi I, MacLennan C, D'Oro U, Saul A, Gerke C
    J Biol Chem, 2014;289(36):24922-35. 2014
  19. Detection of antibodies against Turkey astrovirus in humans.
    Authors: Meliopoulos V, Kayali G, Burnham A, Oshansky C, Thomas P, Gray G, Beck M, Schultz-Cherry S
    PLoS ONE, 2014;9(5):e96934. 2014
  20. Nonfunctional variant 3 factor H binding proteins as meningococcal vaccine candidates.
    Authors: van der Veen, Stijn, Johnson, Steven, Jongerius, Ilse, Malik, Talat, Genovese, Alessia, Santini, Laura, Staunton, David, Ufret-Vincenty, Rafael L, Pickering, Matthew, Lea, Susan M, Tang, Christop
    Infect Immun, 2014;82(3):1157-63. 2014
  21. Dimerization is not a determining factor for functional high affinity human plasminogen binding by the group A streptococcal virulence factor PAM and is mediated by specific residues within the PAM a1a2 domain.
    Authors: Bhattacharya S, Liang Z, Quek A, Ploplis V, Law R, Castellino F
    J Biol Chem, 2014;289(31):21684-93. 2014
  22. Alloimmune activation promotes anti-cancer cytotoxicity after rat liver transplantation.
    Authors: Lacotte S, Oldani G, Slits F, Orci L, Rubbia-Brandt L, Morel P, Mentha G, Toso C
    PLoS ONE, 2014;9(3):e91515. 2014
  23. Cyclooxygenase inhibition abrogates aeroallergen-induced immune tolerance by suppressing prostaglandin I2 receptor signaling.
    Authors: Zhou W, Goleniewska K, Zhang J, Dulek D, Toki S, Lotz M, Newcomb D, Boswell M, Polosukhin V, Milne G, Wu P, Moore M, FitzGerald G, Peebles R
    J Allergy Clin Immunol, 2014;134(3):698-705.e5. 2014
  24. Cationic lipid-formulated DNA vaccine against hepatitis B virus: immunogenicity of MIDGE-Th1 vectors encoding small and large surface antigen in comparison to a licensed protein vaccine.
    Authors: Endmann A, Klunder K, Kapp K, Riede O, Oswald D, Talman E, Schroff M, Kleuss C, Ruiters M, Juhls C
    PLoS ONE, 2014;9(7):e101715. 2014
  25. GPR56 inhibits melanoma growth by internalizing and degrading its ligand TG2.
    Authors: Yang L, Friedland S, Corson N, Xu L
    Cancer Res, 2014;74(4):1022-31. 2014
  26. Netrin-1 controls sympathetic arterial innervation.
    Authors: Brunet I, Gordon E, Han J, Cristofaro B, Broqueres-You D, Liu C, Bouvree K, Zhang J, Del Toro R, Mathivet T, Larrivee B, Jagu J, Pibouin-Fragner L, Pardanaud L, Machado M, Kennedy T, Zhuang Z, Simons M, Levy B, Tessier-Lavigne M, Grenz A, Eltzschig H, Eichmann A
    J Clin Invest, 2014;124(7):3230-40. 2014
  27. Microparticle alpha-2-macroglobulin enhances pro-resolving responses and promotes survival in sepsis.
    Authors: Dalli, Jesmond, Norling, Lucy V, Montero-Melendez, Trinidad, Federici Canova, Donata, Lashin, Hazem, Pavlov, Anton M, Sukhorukov, Gleb B, Hinds, Charles, Perretti, Mauro
    EMBO Mol Med, 2014;6(1):27-42. 2014
  28. Latent TGF-beta binding protein 4 promotes elastic fiber assembly by interacting with fibulin-5.
    Authors: Noda K, Dabovic B, Takagi K, Inoue T, Horiguchi M, Hirai M, Fujikawa Y, Akama T, Kusumoto K, Zilberberg L, Sakai L, Koli K, Naitoh M, von Melchner H, Suzuki S, Rifkin D, Nakamura T
    Proc Natl Acad Sci U S A, 2013;110(8):2852-7. 2013
  29. Shedding of c-Met ectodomain correlates with c-Met expression in non-small cell lung cancer.
    Authors: Fu, Le, Guo, Wei, Liu, Bingshan, Sun, Linlin, Bi, Zhenghon, Zhu, Li, Wang, Xinyan, Liu, Bin, Xie, Qian, Li, Ke
    Biomarkers, 2013;18(2):126-35. 2013
  30. The CD8-derived chemokine XCL1/lymphotactin is a conformation-dependent, broad-spectrum inhibitor of HIV-1.
    Authors: Guzzo C, Fox J, Lin Y, Miao H, Cimbro R, Volkman B, Fauci A, Lusso P
    PLoS Pathog, 2013;9(12):e1003852. 2013
  31. A novel role for insulin resistance in the connection between obesity and postmenopausal breast cancer.
    Authors: Weichhaus M, Broom J, Wahle K, Bermano G
    Int. J. Oncol., 2012;41(2):745-52. 2012
  32. The role of alveolar epithelial cells in initiating and shaping pulmonary immune responses: communication between innate and adaptive immune systems.
    Authors: Chuquimia OD, Petursdottir DH, Rahman MJ, Hartl K, Singh M, Fernandez C
    PLoS ONE, 2012;7(2):e32125. 2012
  33. A Monoclonal Antibody-GDNF Fusion Protein Is Not Neuroprotective and Is Associated with Proliferative Pancreatic Lesions in Parkinsonian Monkeys.
    Authors: Ohshima-Hosoyama S, Simmons HA, Goecks N
    PLoS ONE, 2012;7(6):e39036. 2012
  34. Combined Flt3L/TK gene therapy induces immunological surveillance which mediates an immune response against a surrogate brain tumor neoantigen.
    Authors: King GD, Muhammad AK, Larocque D
    Mol. Ther., 2011;19(10):1793-801. 2011
  35. Adiponectin inhibits leptin signalling via multiple mechanisms to exert protective effects against hepatic fibrosis.
    Authors: Handy JA, Fu PP, Kumar P
    Biochem. J., 2011;440(3):385-95. 2011
  36. Decompensated heart failure is associated with reduced corin levels and decreased cleavage of pro-atrial natriuretic peptide.
    Authors: Ibebuogu UN, Gladysheva IP, Houng AK
    Circ Heart Fail, 2011;4(2):114-20. 2011
  37. Influence of galectin-9/Tim-3 interaction on herpes simplex virus-1 latency.
    Authors: Reddy PB, Sehrawat S, Suryawanshi A
    J. Immunol., 2011;187(11):5745-55. 2011
  38. Apolipoprotein B binding domains: evidence that they are cell-penetrating peptides that efficiently deliver antigenic peptide for cross-presentation of cytotoxic T cells.
    Authors: Sakamoto N, Rosenberg AS,
    J. Immunol., 2011;186(8):5004-11. 2011
  39. PEGylation of interleukin-10 for the mitigation of enhanced pain states.
    Authors: Soderquist RG, Milligan ED, Harrison JA, Chavez RA, Johnson KW, Watkins LR, Mahoney MJ
    J Biomed Mater Res A, 2010;93(3):1169-79. 2010
  40. Macrophage migration inhibitory factor anti-thrombin III complexes are decreased in bladder cancer patient serum: Complex formation as a mechanism of inactivation.
    Authors: Meyer-Siegler KL, Cox J, Leng L, Bucala R, Vera PL
    Cancer Lett., 2010;290(1):49-57. 2010
  41. A novel role for constitutively expressed epithelial-derived chemokines as antibacterial peptides in the intestinal mucosa.
    Authors: Kotarsky K, Sitnik KM, Stenstad H, Kotarsky H, Schmidtchen A, Koslowski M, Wehkamp J, Agace WW
    Mucosal Immunol, 2010;3(1):40-8. 2010
  42. Evaluation of soluble junctional adhesion molecule-A as a biomarker of human brain endothelial barrier breakdown.
    Authors: Haarmann A, Deiss A, Prochaska J, Foerch C, Weksler B, Romero I, Couraud PO, Stoll G, Rieckmann P, Buttmann M
    PLoS ONE, 2010;5(10):e13568. 2010
  43. Enzyme-linked enzyme-binding assay for Pin1 WW domain ligands.
    Authors: Mercedes-Camacho AY, Etzkorn FA,
    Anal. Biochem., 2010;402(1):77-82. 2010
  44. Systemic immunization with CCL27/CTACK modulates immune responses at mucosal sites in mice and macaques.
    Authors: Kraynyak KA, Kutzler MA, Cisper NJ, Khan AS, Draghia-Akli R, Sardesal NY, Lewis MG, Yan J, Weiner DB
    Vaccine, 2010;28(8):1942-51. 2010
  45. A role for calcium-calmodulin in regulating nitric oxide production during skeletal muscle satellite cell activation.
    Authors: Tatsumi R, Wuollet AL, Tabata K, Nishimura S, Tabata S, Mizunoya W, Ikeuchi Y, Allen RE
    Am. J. Physiol., Cell Physiol., 2009;296(4):C922-9. 2009
  46. Endothelin-converting enzyme-2 is increased in Alzheimer's disease and up-regulated by Abeta.
    Authors: Palmer JC, Baig S, Kehoe PG, Love S
    Am. J. Pathol., 2009;175(1):262-70. 2009
  47. Nanobacteria are mineralo fetuin complexes.
    Authors: Raoult D, Drancourt M, Azza S, Nappez C, Guieu R, Rolain JM, Fourquet P, Campagna B, La Scola B, Mege JL, Mansuelle P, Lechevalier E, Berland Y, Gorvel JP, Renesto P
    PLoS Pathog., 2008;4(2):e41. 2008
  48. Up-regulated macrophage migration inhibitory factor protects apoptosis of dermal fibroblasts in patients with systemic sclerosis.
    Authors: Kim JY, Kwok SK, Hur KH, Kim HJ, Kim NS, Yoo SA, Kim WU, Cho CS
    Clin. Exp. Immunol., 2008;152(2):328-35. 2008
  49. Soluble ErbB3 levels in bone marrow and plasma of men with prostate cancer.
    Authors: Lin SH, Lee YC, Choueiri MB, Wen S, Mathew P, Ye X, Do KA, Navone NM, Kim J, Tu SM, Yu-Lee LY, Logothetis CJ
    Clin. Cancer Res., 2008;14(12):3729-36. 2008
  50. Differential gene expression in cultured osteoblasts and bone marrow stromal cells from patients with Paget's disease of bone.
    Authors: Naot D, Bava U, Matthews B, Callon KE, Gamble GD, Black M, Song S, Pitto RP, Cundy T, Cornish J, Reid IR
    J. Bone Miner. Res., 2007;22(2):298-309. 2007
  51. Soluble triggering receptor expressed on myeloid cell-1 is increased in patients with ventilator-associated pneumonia: a preliminary report.
    Authors: Horonenko G, Hoyt JC, Robbins RA, Singarajah CU, Umar A, Pattengill J, Hayden JM
    Chest, 2007;132(1):58-63. 2007
  52. The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production.
    Authors: Powell JM, Sonnenfeld G
    J. Interferon Cytokine Res., 2006;26(1):34-9. 2006
  53. Human cytomegalovirus-specific CD4+-T-cell cytokine response induces fractalkine in endothelial cells.
    Authors: Bolovan-Fritts CA, Trout RN, Spector SA
    J. Virol., 2004;78(23):13173-81. 2004
  54. Selective roles for tumor necrosis factor alpha-converting enzyme/ADAM17 in the shedding of the epidermal growth factor receptor ligand family: the juxtamembrane stalk determines cleavage efficiency.
    Authors: Hinkle CL, Sunnarborg SW, Loiselle D, Parker CE, Stevenson M, Russell WE, Lee DC
    J. Biol. Chem., 2004;279(23):24179-88. 2004
  55. ELR+ CXC chemokines in human milk.
    Authors: Maheshwari A, Christensen RD, Calhoun DA
    Cytokine, 2003;24(3):91-102. 2003
Expand to show all 55 Citations
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DuoSet ELISA Ancillary Reagent Kit 2

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Streptavidin-HRP (set of 5 vials enough for 75 plates total)

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Quantikine Wash Buffer 1

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Clear Polystyrene Microplates, 25 Pack

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Reagent Additive 1 (21 mL)

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Glo Substrate Reagent Pack (Reagent A & B)

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Reagent Diluent Concentrate 1 (21 mL)

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Sample Diluent Concentrate 2 (2X)

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Reagent Diluent Concentrate 3 (5X, 10 x 21 mL)

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Sample Diluent Concentrate 1 (5X, 5 x 21 mL)

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Cell Lysis Buffer 1 (1 x 21 mL)

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Black Polystyrene Microplates, 25 Pack

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Cell Lysis Buffer 5 (1 x 21 mL)

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Cell Lysis Buffer 3 Concentrate (5X, 1 x 21 mL)

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Average Rating: 4.6 (Based on 13 reviews)

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  Excellent
  Anonymous 09/07/2017
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Other Experimental Details

Other Experimental DetailsOur lab does many ELISAs. These substrate solutions are excellent, affordable, and easy to use.
 Substrate Reagent Pack (8 vials Color A, 8 vials Color B) DY999
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  Anonymous 07/19/2017
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: Substrate Reagent Pack (8 vials Color A, 8 vials Color B) [DY999]
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  Anonymous 07/17/2017
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  Anonymous 07/14/2017
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 Substrate Reagent Pack (8 vials Color A, 8 vials Color B) DY999
: Substrate Reagent Pack (8 vials Color A, 8 vials Color B) [DY999]
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  Anonymous 07/06/2017
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  Anonymous 06/15/2017
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 Substrate Reagent Pack (8 vials Color A, 8 vials Color B) DY999
: Substrate Reagent Pack (8 vials Color A, 8 vials Color B) [DY999]

Other Experimental Details

Other Experimental DetailsReally Easy to use.After the SA-HRP detection step for ELISA, wash wells 4 times with wash buffer, and add the Substrate and incubate for 20 mins in the dark. Stop the reaction using 0.2 M sulfuric acid. Read the plate at 450nm
  Excellent
  Anonymous 05/03/2017
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  Excellent
  Anonymous 04/19/2017
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 Substrate Reagent Pack (8 vials Color A, 8 vials Color B) DY999
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  Anonymous 03/29/2017
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: Substrate Reagent Pack (8 vials Color A, 8 vials Color B) [DY999]
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  Anonymous 11/28/2016
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  Anonymous 09/01/2016
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: Substrate Reagent Pack (8 vials Color A, 8 vials Color B) [DY999]
 Substrate Reagent Pack (8 vials Color A, 8 vials Color B) dy999
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